Stability and Reactivity of 2-Nitrosoamino-3,8-dimethylimidazo[4,5-f]quinoxaline

• Published in: Chemical research in toxicology Vol. 19; no. 2; pp. 325 - 333
• Main Authors: Lakshmi, Vijaya M, Hsu, Fong Fu, Schut, Herman A. J, Zenser, Terry V
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.02.2006
• Subjects: Animals; Chromatography, High Pressure Liquid; DNA - chemistry; DNA - drug effects; DNA - isolation & purification; DNA Adducts - chemistry; DNA Adducts - drug effects; Dose-Response Relationship, Drug; Hydrogen-Ion Concentration; Imidazoles - chemistry; Imidazoles - pharmacology; Liver - chemistry; Liver - drug effects; Male; Mice; Mice, Inbred C57BL; Nitrosamines - chemistry; Nitrosamines - pharmacology; Quinoxalines - administration & dosage; Quinoxalines - chemistry; Quinoxalines - pharmacology; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; Time Factors
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/tx050305x

2-Nitrosoamino-3,8-dimethylimidazo[4,5-f]quinoxaline (N-NO-MeIQx) is a nitrosation product of the food carcinogen 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and is proposed to form in vivo under inflammatory conditions. This study evaluated the stability and reactivity of N-NO-MeIQx to assess its possible role in the initiation of colon cancer by MeIQx. 14C-N-NO-MeIQx (4 μM) was incubated for 4 h over a range of pH values, and its stability was monitored by HPLC. At pH values from pH 7.4 to 9.0, N-NO-MeIQx was very stable with no detectable change observed. Glutathione (1 mM) did not alter stability at pH 7.4. As the pH decreased, this nitrosamine was less stable with only 48 ± 1% remaining at pH 5.5 and none remaining at pH 3.5 or 2.0. Major products identified by electrospray ionization mass spectrometry were 3,8-dimethylimidazo[4,5-f]quinoxaline and 2-hydroxy-3,8-dimethylimidazo[4,5-f]quinoxaline. MeIQx was a minor product. At pH 2.0, the t 1/2 for N-NO-MeIQx was reduced from 2.1 ± 0.2 to 1.2 ± 0.1 min with 10 mM NaN3. This effect of azide was due to the formation of 2-azido-MeIQx. The binding of 14C-N-NO-MeIQx to DNA increased with decreasing pH. The 10-fold increase in binding observed at pH 2.0 as compared to pH 5.5 was completely inhibited by 10 mM NaN3 due to 2-azido-MeIQx formation. The reactivity of N-NO-MeIQx was compared to N-OH-MeIQx by evaluating adduct formation with 2‘-deoxyguanosine 3‘-monophosphate (dGp) by 32P-postlabeling. N-OH-MeIQx formed a single major adduct, N-(deoxyguanosin-8-yl)-MeIQx (dG-C8-MeIQx). Incubation of N-NO-MeIQx under inflammatory conditions (pH 5.5 ± HOCl) produced dG-C8-MeIQx along with 4−6 other adducts. dG-C8-MeIQx formation increased in the presence of HOCl. Liver from a MeIQx-treated mouse contained dG-C8-MeIQx and two other adducts detected with N-NO-MeIQx but not N-OH-MeIQx. These results suggest that N-NO-MeIQx could be genotoxic, is activated by conditions that mediate inflammatory responses, and is a possible cancer risk factor for individuals with inflammation of the colon.